CN102341706A - Method for Detecting Prozone Phenomenon, Analysis Method, Device for Detecting Prozone Phenomenon, and Analysis Device - Google Patents
Method for Detecting Prozone Phenomenon, Analysis Method, Device for Detecting Prozone Phenomenon, and Analysis Device Download PDFInfo
- Publication number
- CN102341706A CN102341706A CN2010800098989A CN201080009898A CN102341706A CN 102341706 A CN102341706 A CN 102341706A CN 2010800098989 A CN2010800098989 A CN 2010800098989A CN 201080009898 A CN201080009898 A CN 201080009898A CN 102341706 A CN102341706 A CN 102341706A
- Authority
- CN
- China
- Prior art keywords
- prozone phenomenon
- test section
- sample
- target composition
- prozone
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/557—Immunoassay; Biospecific binding assay; Materials therefor using kinetic measurement, i.e. time rate of progress of an antigen-antibody interaction
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
- G01N33/537—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody
- G01N33/538—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with separation of immune complex from unbound antigen or antibody by sorbent column, particles or resin strip, i.e. sorbent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
- G01N33/54387—Immunochromatographic test strips
- G01N33/54388—Immunochromatographic test strips based on lateral flow
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Provided is a prozone phenomenon detecting method, by which generation of a prozone phenomenon can be detected even when a conventional specimen analysis tool is used, and examinations using an immunochromatography method and the like can be performed efficiently. In the method, a specimen analysis tool containing substances that specifically bind to a target component contained in a sample is used. The specimen analysis tool is obtained by arranging a sample supplying portion, a reagent portion, and a detection portion on the porous base material from upstream to downstream in a sample moving direction in this order. The reagent portion contains a labeled substance that specifically binds to the target component. The detection portion contains an immobilized substance that specifically binds to the target component. The target component is detected by detecting a complex of the target component, the labeled substance, and the immobilized substance through detection of a label of the labeled substance in the detection portion. The method includes at least one of the following processes A and B: the process A: a process in which detection results obtained in the detection portion are plotted along the sample moving direction, and generation of a prozone phenomenon is detected on the basis of a position of a peak in plots thus obtained; and the process B: a process in which the label is detected at two or more different time points in the detection portion, and generation of a prozone phenomenon is detected on the basis of a magnitude relationship between two or more detection results thus obtained.
Description
Technical field
The present invention relates to prozone phenomenon detection method, analytical approach, prozone phenomenon pick-up unit and analytical equipment.
Background technology
In recent years, in the diagnosis of medical field, be used for pathogen such as bacterial detection, virus and be used to judge whether that conceived analyte analysis tool is popularized.In the said analyte analysis tool, therefore the analyte analysis tool that utilizes immunochromatography obtains widespread use owing to can easy and promptly detect.The said detection principle of the analyte analysis tool of immunochromatography of utilizing for example is described below.That is, at first prepare to form and be fixed with the analyte analysis tool of immobilized antibody at the test section of said porous substrate by porous substrate.To wherein adding sample (determinand) and with the marking antibody of colored particles mark.When the antigen that exists in the said sample as the analytic target composition, said marking antibody and said immobilized antibody are situated between and form compound by said antigen, and the test section on the porous substrate is painted through said colored particles.As said mark, except colored particles, also comprise enzyme and the combination etc. of painted substrate through enzyme reaction.Said test section is generally wire.Confirm under the painted situation, thereby judge in the sample and exist the analytic target composition to be judged to be the positive, if not painted then judge do not have the analytic target composition in the sample and be judged to be feminine gender.In said immunochromatography, also carried out following trial:, the analytic target composition in the sample is carried out half-quantitative detection (for example with reference to patent documentation 1) through setting a plurality of said test sections or passing through the painted of the stage of recognition property.
In the detection of the analytic target composition that uses immunochromatography, the prozone phenomenon that the concentration of analytic target composition is produced when high becomes problem.Prozone phenomenon is meant, do not exist or the phenomenon of low concentration though the analytic target composition in the actual samples is judged as on to be high concentration apparent.Fig. 5 is the figure of the relation of the absorbance (coloring degree) of antigen concentration and test section in the expression immunochromatography.Absorbance raises along with antigen concentration and rises, but when reach finite concentration when above generation can not get the such phenomenon of absorbance (not painted).Therefore, detect absorbance X when for example antigen concentration is for Z1, but under the concentration Z2 of area with high mercury, only detect the absorbance of same degree owing to produce prozone phenomenon.This phenomenon is to result from when the analytic target composition in sample during excessive the existence, and said immobilized antibody quilt and the embedding of the nonreactive analytic target composition of marking antibody can't be captured by said immobilized antibody with the analytic target composition of marking antibody response.Therefore area with high mercury is apparent sometimes negative (false negative), be difficult to difference in analyzing with the situation of real feminine gender.When the false negative due to suspection is prozone phenomenon, need the suitable dilution of sample be checked again.On the other hand; In order to detect the generation of prozone phenomenon; Following trial has been proposed: as the immobilized antibody of analyte analysis tool, use the different multiple antibody (for example with reference to patent documentation 2) of compatibility, or in a plurality of test sections the stage sexually revise the amount (for example with reference to patent documentation 3) of antibody.
The prior art document
Patent documentation
Patent documentation 1: japanese kokai publication hei 8-278305 communique
Patent documentation 2:WO2003/014740
Patent documentation 3: No. 3644780 communique of Jap.P..
Summary of the invention
The problem that invention will solve
Yet, the structure of analyte analysis tool is complicated.In addition, can't detect the generation of prozone phenomenon before in the zone in the downstream of a plurality of test sections, finishing reaction, checking efficiency possibly reduce.These problems be not only be present in utilized with determinand in contained analytic target composition be that antigen carries out in the immunoreactive immune analysis method of the antibody that specificity combines, use with determinand in all possibly take place in all analytical approachs of the material (specificity junction mixture matter) that combines of contained analytic target composition specificity.
Therefore the object of the present invention is to provide prozone phenomenon detection method, analytical approach, prozone phenomenon pick-up unit and analytical equipment; Even said prozone phenomenon detection method uses existing analyte analysis tool also can detect the generation of prozone phenomenon simply, and can efficiently use the inspection of immunochromatography etc.
The scheme that is used to deal with problems
To achieve these goals, the detection method of prozone phenomenon of the present invention is characterized in that,
Use contain with sample in the analyte analysis tool of the material that combines of analytic target composition specificity,
Said analyte analysis tool is for disposing the analyte analysis tool of sample supply portion, reagent portion and test section downstream from the upper reaches of the moving direction of said sample on porous substrate,
The marking material that combines with said analytic target composition specificity is contained in said reagent portion,
Said test section contains the immobilization material that combines with said analytic target composition specificity,
Through at said test section the compound of said analytic target composition, said marking material and said immobilization material being detected the said mark of said marking material, thereby said analytic target composition is able to detect,
Said method comprises at least a in following A method and the B method,
A method: at said test section, testing result is mapped, by the generation of the position probing prozone phenomenon at the peak of said figure along the moving direction of said sample;
B method:, change time and implement the detection of 2 the above marks, detect the generation of prozone phenomenon by the magnitude relationship of 2 above testing results that obtain at said test section.
Analytical approach of the present invention is characterized in that, comprises analysis procedure and prozone phenomenon and detects operation,
Said analysis procedure is used analyte analysis tool and is implemented,
Said analyte analysis tool is for disposing the analyte analysis tool of sample supply portion, reagent portion and test section downstream from the upper reaches of the moving direction of said sample on porous substrate,
The marking material that combines with said analytic target composition specificity is contained in said reagent portion,
Said test section contains the immobilization material that combines with said analytic target composition specificity; Through the compound of said analytic target composition, said marking material and said immobilization material being detected the said mark of said marking material at said test section; Thereby said analytic target composition is able to detect
Said prozone phenomenon detects operation and implements through said prozone phenomenon detection method of the present invention.
Prozone phenomenon pick-up unit of the present invention is characterized in that, it is used for said prozone phenomenon detection method of the present invention,
Comprise the testing result that is used to obtain said test section obtain the unit and
At least a in following A unit and the B unit,
The A unit: the detecting unit of the generation of the position probing prozone phenomenon at peak that get, said testing result by mapping along the moving direction of said sample,
B unit: change time and implement the detection of 2 the above marks, detect the detecting unit of the generation of prozone phenomenon by the magnitude relationship of 2 above testing results that obtain.
Analytical equipment of the present invention is characterized in that, it is used for said analytical approach of the present invention,
Comprise analytic unit and prozone phenomenon detecting unit,
Said analytic unit is through detecting the said mark of said marking material to the compound of said analytic target composition, said marking material and said immobilization material at the said test section of said analyte analysis tool; Thereby detect said analytic target composition
Said prozone phenomenon detecting unit is said prozone phenomenon pick-up unit of the present invention.
The effect of invention
According to the present invention,, and can efficiently use the inspection that utilizes immunochromatography that specificity junction mixture matter analyzes etc. even use existing analyte analysis tool also can detect the generation of prozone phenomenon simply.Therefore, we can say that the present invention is exceedingly useful in analysis, clinical field etc.
Description of drawings
Fig. 1 be the test samples among the embodiment 1 reflected light and detection figure.(A) of Fig. 1 is the result of the sample of 60mg/100mL for CRP concentration, and (B) of Fig. 1 is the result of the sample of 6.5mg/100mL for CRP concentration.
Fig. 2 be 2nd test section of expression among the embodiment 1 the zone, downstream, the reflectivity (R after 10 minutes
10) and poor (the Δ R of reflectivity
5-10) the chart of relation.
(A) of Fig. 3 is the planimetric map of an example of the analyte analysis tool that uses among the present invention of expression, and (B) of Fig. 3 is the process flow diagram of an example of expression prozone phenomenon detection method of the present invention.
The planimetric map of other example of the analyte analysis tool that (A) of Fig. 4 uses among the present invention for expression.(B) of Fig. 4 is the I-I direction cut-open view of the analyte analysis tool shown in (A) of presentation graphs 4.(C) of Fig. 4 is the planimetric map of in housing, taking in the analyte analysis chip of said analyte analysis tool.
Fig. 5 is the key diagram that is used to explain prozone phenomenon.
Description of reference numerals
1 sample supply portion
2 reagent portions
10,20 analyte analysis tools
11 porous substrates
21 infrabasal plates
Porous body is used in 22 detections
23,24 samples are used porous body
25 marking antibody are used porous body
Porous body is used in 26 absorptions
27 test sections
30 analyte analysis chips
31 housings
L1 the 1st test section
L2 the 2nd test section
Test section is used in the C contrast
Embodiment
In the present invention; " prozone phenomenon " is meant;, the analytic target composition in the actual samples do not have or the phenomenon of low concentration the phenomenon of appreciable prozone phenomenon appearance during prozone phenomenon and the institutes such as biochemical field beyond the antigen-antibody reaction that comprises antigen-antibody reaction responds though being high concentration but be judged as on apparent.
In the present invention; In comprising the prozone phenomenon detection method of said B method; Preferably with reference to make in advance, the magnitude relationship of said 2 above testing results has been carried out related judgment standard with the relation of the generation of said prozone phenomenon, detect the generation of said prozone phenomenon.
And then the magnitude relationship of said testing result is preferably at least a in the ratio of difference and 2 above testing results of 2 above testing results.
When in prozone phenomenon detection method of the present invention and analytical approach, comprising said A method and when in prozone phenomenon pick-up unit and analytical equipment, comprising said A unit; The peak position of said figure is positioned under the situation in downstream of moving direction of said sample, can be judged as and produce prozone phenomenon.
In prozone phenomenon detection method of the present invention, analytical approach, prozone phenomenon pick-up unit and analytical equipment, can't be judged as under the situation that has produced prozone phenomenon, and then preferably carry out said B method through said A method.
In prozone phenomenon detection method of the present invention, analytical approach, prozone phenomenon pick-up unit and analytical equipment; 2 above test sections that said test section disposes for the moving direction along sample; In said 2 above test sections; The test section of the upstream side of the moving direction of preferred sample is the test section that is used for the check and analysis object component, and the test section in downstream is the test section that is used to detect prozone phenomenon.
In prozone phenomenon detection method of the present invention, analytical approach, prozone phenomenon pick-up unit and analytical equipment; At the test section that is used for detecting prozone phenomenon, preferably the testing result in the zone in the downstream of the moving direction through said sample detects prozone phenomenon.
In prozone phenomenon detection method of the present invention, analytical approach, prozone phenomenon pick-up unit and analytical equipment, said testing result is preferably optical signalling.
In analytical approach of the present invention, when detecting the generation of said prozone phenomenon, preferably be judged to be false negative.
In prozone phenomenon detection method of the present invention, analytical approach and analytical equipment, be under the situation of antigen at said analytic target composition, said marking material and said immobilization material are that marking antibody and immobilized antibody get final product.In addition, be under the situation of antibody at said analytic target composition, said marking material and said immobilization material are that marking antibody and immobilized antigen get final product.When the analytic target composition was antibody, the antibody of analytic target composition was connected with immobilized antigen, and the front end of the Fab of marking antibody is connected with the Fc zone of the antibody of analytic target composition.
For example the present invention is described in detail below.
In the present invention, as the sample that contains the analytic target composition (determinand), be preferably aqueous sample, but be not limited to this.In the present invention, when said sample is solid, shaped, for example can be dissolved or dispersed in the liquid such as damping fluid and processes solution.Said damping fluid is not special to be limited, and can enumerate out for example Tris damping fluid, phosphate buffer, acetate buffer solution, borate buffer etc.In the present invention; Said sample for example can enumerate out nasal cavity aspirated liquid, nasal cavity cleansing solution, nasal cavity wiping liquid, nasal mucus, pharyngeal wiping liquid, GARG, saliva, whole blood, serum, blood plasma, just, organism sample such as urine, marrow liquid; Food such as foods such as animal and plant, processed food; River in the environmental test etc., not special the qualification.In the present invention, when the analytic target composition is antigen or antibody (below be sometimes referred to as " antigen etc. "), for example can uses and extract the sample that antigen or antibody prepares through extract etc. by said sample and analyze.Said extract is not special to be limited, and for example can enumerate out said damping fluid etc.In addition, can suitably add surfactant, stabilization agent, antiseptic etc. in the said extract.
In the present invention, the sample to analytic target composition, application has no qualification.The present invention is preferred in the detection of following substances, and said material for example is transferrins in granulocyte elastase in the C reactive protein (CRP), HbA1c, TSH, FT3, FT4, hCG, HBs antigen, HBc antibody, HCV antibody, TY antigen, antistreptolysin O (ASO) (ASO), IV Collagen Type VI, matrix metalloproteinase (MMP-3), PIVAK-II, α 1 microglobulin, β 1 microglobulin, amyloid A (SAA), elastoser 1, alkaline alpha-fetoprotein (BFP), candida antigens, cervical mucus in the blood, digoxin, cysteine proteinase inhibitor C, the XIII factor, the urine, syphilis, hyaluronic acid, fibrin monomer compound (SFMC), the pseudohemophilia factor (VIII factor appearance antigen), Protein S, rheumatoid factor (RF), IgD, α 1 acidoglycoprotein (α 1AG), alpha1 Anti-trypsin (α 1AT), alpha2 Macroglobulin, albumin (Alb), CER (Cp), haptoglobin (Hp), prealbumin, RBP ELISA (RBP), β 1C/ β 1A globulin (C3), β 1E globulin (C4), IgA, IgG, IgM, beta Lipoprotein (β-LP), apolipoprotein A-1, apolipoprotein A-1 I, apolipoprotein B, apoC-II, apoC-III, apo E, transferrins (Tf), urinary albumin, plasminogen (PLG), lipoprotein (a) (LP (a)) etc.
In the present invention, when using said marking antibody as said marking material, said marking antibody is not special to be limited.Said marking antibody for example can be the antibody that is combined with painted insoluble carrier granular, also can be the antibody that is combined with enzyme.
In the present invention; Said painted insoluble carrier granular is not special to be limited, and can enumerate out for example colored latex particle, metal colloid particles, painted poly methyl methacrylate particle, painted PLA particle, painted porous glass particle, painted silica dioxide granule, painted agarose particle, painted glucan particles etc.Said colored latex particle is not special to be limited, and for example can enumerate out blue latex particle, red latex particle etc.Said metal colloid particles is not special to be limited, and for example can enumerate out gold colloid particles, platinum colloidal solid etc.The mean grain size of said painted insoluble carrier granular is not special to be limited; Under the situation of said colored latex particle; Particle diameter is for example in the scope of 0.05 μ m～5 μ m, and the scope of preferred 0.1 μ m～1 μ m is under the situation of said metal colloid particles; Particle diameter is for example in the scope of 2nm～100nm, the scope of preferred 10nm～50nm.
In the present invention, said enzyme uses is for example to react and make the painted enzyme of substrate, can enumerate out for example peroxidase, alkaline phosphatase, beta-D-galactosidase etc.
In the present invention, as long as said substrate does not limit also painted with said enzyme reaction especially.Said substrate for example can enumerate out 2; 2 '-azine group-two (3-ethyl benzo thiazole phenanthrolines-6-sulfonic acid) (ABTS), 3; 3 '; 5,5 '-tetramethyl benzidine (TMB), diaminobenzidine (DAB), 5-bromo-4-chloro-3-indoles phosphoric acid (BCIP), 4-methyl umbelliferone base-β-D-galactoside (4MUG), 3-(2 '-spiral diamantane)-4-methoxyl-4-(3 "-β-D-galactopyranose base) phenyl-1,2-dioxetane (AMGPD) etc.
In the present invention, the antibody of said marking antibody is as long as combines with antigen or antibody specificity as the analytic target composition, and not qualification especially can be enumerated out the antibody that for example is directed against said various antigens etc.Said antibody for example can be the antibody that comes from biosome, also can be the antibody of synthetic.The said antibody that comes from biosome can be enumerated out for example immunoglobulin (Ig) (Ig), antibody fragment, chimeric antibody etc.Said immunoglobulin (Ig) can be enumerated out for example IgG, IgA, IgM, IgD, IgE and IgY etc.Said antibody fragment can be enumerated out Fab, Fab ', F (ab ') 2 etc.Said chimeric antibody can be enumerated out for example humanized antibodies etc.In addition, in the present invention, said antibody for example can be prepared through existing known method by the serum that comes from mouse, rabbit, goat, sheep, chicken etc., perhaps also can utilize commercially available various antibody, not special restriction.And then in the present invention, said antibody can use any one in polyclonal antibody and the monoclonal antibody, can be according to suitably settings such as said antigens.The antibody of said synthetic can be enumerated out for example affine body (affibody) etc.Affine body (affibody) is a kind of of artificial antibody, in order to combine with the target substance specificity from the storehouse screening and obtain.It is littler that Affibody and antibody are compared size, in addition heat, alkali etc. had patience.In the present invention, the antibody of said immobilized antibody is the specific antibody to said antigen as the analytic target composition, for example can be identical with said marking antibody, the antibody that can combine with antigen.The kind of said immobilized antibody, preparation method are identical with the preparation method of for example said marking antibody.
In the present invention, the antigen of said immobilized antigen is not special to be limited, and for the specific antigen to said antibody as the analytic target composition, can enumerate out for example above-mentioned various antigens etc.In the present invention, the preparation method of said immobilized antigen can enumerate out and for example have known method etc. now, not special restriction.Sometimes said immobilized antibody or said immobilized antigen are called " immobilized antibody etc. " below.
Next, enumerate immune analysis method as an example, prozone phenomenon detection method of the present invention is described.But prozone phenomenon detection method of the present invention is not limited to following example.
In the planimetric map of Fig. 3 (A), illustrate an example of the said analyte analysis tool that uses in the immune analysis method.As shown in the figure, in this analyte analysis tool 10, the upper reaches from the moving direction (arrow) of said sample on porous substrate 11 have sample supply portion 1, reagent portion the 2, the 1st test section (L1), the 2nd test section (L2) and contrast downstream with test section (C).In this analyte analysis tool 10, comprising said the 1st test section (L1), said the 2nd test section (L2) and said contrast is said test section with the zone of test section (C).Said reagent portion's 2 underlinedization of load antibody (the for example anti-CRP antibody of blue latex mark).Said the 1st test section (L1) is fixed with anti-CRP antibody for being used to detect the test section of the antigen (for example CRP) as the analytic target composition.Said the 2nd test section (L2) is the test section that is used to detect prozone phenomenon, is fixed with for example anti-CRP antibody.Be fixed with the same antibody of same amount among said L1 and the L2.Said contrast is the control test portion that is fixed with for example anti-IgG antibody with test section (C).
In the present invention, as long as said porous substrate 11 has the capillary porous structure of performance, not special the qualification.Said porous substrate 11 can be enumerated out for example cellulose derivative films such as cellulose membrane, acetyl cellulose, NC Nitroncellulose, glass filter, filter paper etc.In the present invention, the shape of said porous substrate is not special to be limited, and can enumerate out for example rectangle, circle etc.In the present invention, the not special restriction of said porous substrate size for example can be according to suitable settings the such as specification of analytical equipment.
Be formed with the test section of three wire of extending in this routine analyte analysis tool, but the present invention is not limited only to this at the Width of said porous substrate.The number of said test section can freely be set in 1～10 scope, and the shape of test section can also be enumerated out for example rectangle, circle etc. except that wire in addition.
Next, based on (A) of Fig. 3 and (B), an example of the prozone phenomenon detection method in the immune analysis method that uses this routine analyte analysis tool is described.
(prozone phenomenon of A method detects)
At first gather serum (sample) (step S1).Next, said sample is added drop-wise to said sample supply 1 (the step S2) of portion.Through said dropping, said sample is supplied to said reagent portion 2, and dissolves the said marking antibody of 2 loads of said reagent portion.When containing the antigen as the analytic target composition in the said sample, combine, form antigen-marking antibodies body through said marking antibody of antigen-antibody reaction and said antigen.Said then antigen-marking antibodies body launches in said porous substrate, combines with the said immobilized antibody of said test section and forms compound.Because form said compound, said the 1st test section (L1), said the 2nd test section (L2) and said contrast develop the color through said mark with test section (C).
Next, be used to represent that in time (t1) detection said test section has or not the reflected light of colour developing and colour developing degree.Particularly, at first prepare following figure, said figure is (the step S3) that along the moving direction (arrow) of said sample said catoptrical testing result is mapped and got at said test section.Next, detection is positioned at said contrast with near the paddy type peak the coordinate of test section (C).Under the situation that can't detect this paddy type peak, can think that said sample and said marking antibody normally do not move, and therefore can't measure it.Next, near the coordinate of said the 1st test section (L1) and said the 2nd test section (L2), detect 2 paddy type peaks.The type peak, mountain on border that will be positioned at said 2 paddy type peaks here, is as the border of said the 1st test section (L1) and said the 2nd test section (L2).With the coordinate on this border as the terminal coordinate of the upstream side of said the 2nd test section (L2), on the other hand will from the coordinate on this border downstream side add setting coordinate as the terminal coordinate in the downstream of said the 2nd test section (L2).Then, connect the figure of the detected value of the terminal coordinate in figure and the said downstream of detected value of the terminal coordinate of said upstream side with line segment, with said line segment and said paddy type figure institute area surrounded as the whole peak of said the 2nd test section (L2).In the present invention, detect the generation of prozone phenomenon by the shape (peak position) of the figure of test section.Preceding half (upstream side) that preferably be positioned at the whole peak of said the 2nd test section (L2) through said peak position still is positioned at later half (downstream), judges whether to produce prozone phenomenon.Particularly, judge at first whether said peak position is positioned at later half (downstream) (the step S4) at the whole peak of said the 2nd test section (L2).Be not positioned under the situation (denying) in downstream, as after the prozone phenomenon of the B method stated detect the generation of prozone phenomenon detecting.Be positioned under the situation (being) in downstream, be judged as and produced prozone phenomenon (step S5).For example in said the 2nd test section (L2),, said peak is divided into " preceding half " and " later half " for example further with the datum line separated into two parts of said peak with the mid point that passes through said line segment.Then, obtain the minimum value (preceding half: L2f-min, later half: L2l-min) of the figure of the detected value in each scope.Then if L2f-min >=L2l-min then is judged as and has produced prozone phenomenon.
In above-mentioned, use the datum line of the mid point through said line segment said peak separated into two parts is judged whether said peak position is positioned at later half (downstream) at the whole peak of said the 2nd test section (L2), but be not to be defined in this.Also can be with the line of the for example point in the downstream 1/3rd through said line segment etc. as said datum line, and be that the border defines upstream side and downstream with this datum line.Can be based on the distance of size, sample supply portion and the test section of the detection characteristic of for example analytic target composition, test section, measure decision such as needed time where datum line be located at.
In addition, under the situation about when having a plurality of test section, compare, the test section peak height in downstream is higher, also can be judged as and produce prozone phenomenon at peak height to the test section in the test section of upstream side and downstream.
Think that one of the reason of prozone phenomenon is; In the sample antigen with can be excessive the existence more than the amount of the marking antibody response of institute of reagent portion load; Therefore marking antibody and unreacted antigen flow out to test section, and with the antibody response that is fixed on test section.In this case, marking antibody and unreacted antigen block the antibody that is fixed on test section, therefore after this flow through that come and the antigen marking antibody response can not with the antibody response that is fixed on test section, colour developing shoals.This phenomenon seems more remarkable at the upstream side of test section than the downstream.Therefore, if there is the peak in test section later half then can be judged as and produced prozone phenomenon.
Example is illustrated in the example that said the 2nd test section (L2) detects prozone phenomenon in above-mentioned, but also can detect at said the 1st test section (L1).In addition, analyte analysis tool has under the situation of 3 above test sections, also can detect at other any test section.
(prozone phenomenon of B method detects)
, the prozone phenomenon of said A method is judged as the situation that does not produce prozone phenomenon in detecting; Next; To being result's mapping of the detection of reflected light obtained in advance the time (t2) in (t1) the preceding moment in elapsed time, same this reflected light is used to be illustrated in said test section and has or not colour developing and colour developing degree (step S4-1) during with the time (t1).Then, for example by detecting prozone phenomenon from time (t2) of the reflectivity of said the 2nd test section (L2) variable quantity through the time (t1).At this moment, can detect prozone phenomenon through the whole peak of said the 2nd test section (L2), but preference as again with the coordinate of the mid point through said line segment with said peak separated into two parts, and the zone detection prozone phenomenon in the downstream through said the 2nd test section (L2).At first, at time (t1), the testing result to the zone in the downstream of said the 2nd test section (L2) is converted into the ratio (%) from said line segment with count value, adds up to obtain reflectivity R
T1(%).Likewise,, detect the reflected light of said test section, obtain reflectivity R at time (t2)
T2(%).From the not special restriction of the time of detecting beginning to the said time (t1), for example in 5～20 minutes scope, preferably in 5～15 minutes scope, more preferably 8～12 minutes scope.The also not special restriction of the mistiming (t1-t2) of said time (t1) and said time (t2), for example 2～8 minutes scope, preferred 3～7 minutes scope, more preferably 4～6 minutes scope.Poor (Δ R by 2 reflectivity that obtain
T2-t1(%)=R
T2-R
T1) detect the generation of prozone phenomenon.Calculating of said catoptrical detection and said reflectivity is not limited to said time (t1) and said time (t2), also can carry out more than 3 times.In addition, also can be for example every carry out said catoptrical detection through one minute, the testing result that makes the said time (t2) is the mean value of said time (t2)-1 for example minute, said time (t2), minute these 3 testing results of said time (t2)+1.
Here, preferably with reference to make in advance, the difference of said reflectivity has been carried out the generation that related judgment standard detects said prozone phenomenon with the relation of the generation of prozone phenomenon.Said judgment standard for example can enumerate out the relation mapping of the generation of the difference of said reflectivity and said prozone phenomenon and chart in the table etc. of relation of generation of difference and said prozone phenomenon of datum line, the said reflectivity of expression.Use to the relation mapping of the generation of the difference of said reflectivity and said prozone phenomenon and chart in the situation of datum line under, also can through least square method make the boundary line and with it as datum line.Particularly, at first judge poor (the Δ R of said reflectivity
T2-t1) whether be lower than datum line (step S4-2).(be) to be judged as to have produced prozone phenomenon (step S5) when being lower than datum line.(deny) to be judged as not produce prozone phenomenon (step S6) when being not less than datum line.
In the present invention, can be detected the generation of said prozone phenomenon by the ratio of the difference of said reflectivity and said reflectivity, perhaps the ratio with said reflectivity replaces the difference of said reflectivity to detect the generation of said prozone phenomenon; Also can detect the generation of said prozone phenomenon by the difference of said reflectivity with than the magnitude relationship of said reflectivity in addition.In addition, in this example, use reflectivity to come certification mark, but also can use transmissivity, absorbance etc. as optical signalling.
Said analytic target composition can be oxidized situation under; In the detection of the generation of said prozone phenomenon; Also can utilize magnitude relationship and the magnitude relationship of oxidation current value of the caused reflectivity of colour developing of the marking antibody of above-mentioned that kind to detect, perhaps utilize the magnitude relationship of oxidation current value to replace the magnitude relationship of the caused reflectivity of colour developing of the marking antibody of above-mentioned that kind to detect.In this case, uses such as said marking antibody be antibody or the antigen that is combined with oxidoreducing enzyme, and at said test section configuration electron accepter and electrode (negative electrode and anode).Said oxidoreducing enzyme so long as enzyme that can the said analytic target composition of oxidation get final product.When using such oxidoreducing enzyme, for example can detect the generation of prozone phenomenon with being described below.That is, in said the 2nd test section (L2), through the catalytic reaction of said oxidoreducing enzyme, said analytic target composition is oxidized, and said electron accepter is reduced simultaneously.Then through electrochemical method with the oxidation once more of the said electron accepter that is reduced.Through this once more the oxidation current value that gets of oxidation become component with said analytic target be corresponding, therefore can carry out quantitatively said analytic target composition indirectly through measuring said electric current.Said electrode is not special to be limited, and can enumerate out for example gold electrode, carbon resistance rod, silver electrode etc.In addition, in said analyte analysis tool, said electrode is optional component parts.Said electrode for example can be configured in said test section when using said analyte analysis tool.
In above-mentioned; The mode of further implementing the prozone phenomenon detection of said B method when not producing prozone phenomenon in the prozone phenomenon detection of said A method, being judged as is illustrated; In the present invention, also can in said A method and said B method, adopt said A method separately or adopt said B method to carry out the prozone phenomenon detection separately.
Next, analytical approach of the present invention is described.As previously mentioned, analytical approach of the present invention comprises analysis procedure and prozone phenomenon detection operation.Said analysis procedure uses analyte analysis tool to implement.Used identical in said analyte analysis tool and the said prozone phenomenon detection method of the present invention.Said prozone phenomenon detects operation and implements through said prozone phenomenon detection method of the present invention.
In analytical approach of the present invention, said prozone phenomenon detects when detecting the generation of prozone phenomenon in the operation, and it is preferred that the situation that the analysis of the analytic target composition in the sample can't accurately be judged is judged to be false negative.Through being judged to be false negative, thereby distinguish, and can carry out more definite analysis with feminine gender.In addition, owing to detected the generation of prozone phenomenon, therefore will not analyze once more after the sample dilution.
Next, prozone phenomenon pick-up unit of the present invention is described.As previously mentioned; Prozone phenomenon pick-up unit of the present invention; It is characterized in that; It is the prozone phenomenon pick-up unit that is used for said prozone phenomenon detection method of the present invention, and what this prozone phenomenon pick-up unit comprised the testing result that is used for obtaining said test section obtains at least a of unit and following A unit and B unit.Here; The A unit is: by the detecting unit along the generation of the position probing prozone phenomenon at the moving direction of said sample mapping peak that get, said testing result; The B unit is: the change time is implemented the detection of 2 the above marks, is detected the detecting unit of the generation of prozone phenomenon by the magnitude relationship of 2 above testing results that obtain.
In said prozone phenomenon detection method of the present invention, under the situation of detection of reflected light as said, the said unit of obtaining has for example light source portion and light accepting part.Shine irradiates light by said light source portion comprising on said the 1st test section (L1) of said analyte analysis tool 10, said the 2nd test section (L2) and the porous substrate of said contrast with test section (C), and through said light accepting part detection of reflected light.Said light source portion is by for example light emitting diode (LED), semiconductor laser diode formations such as (LD).Said light accepting part is by for example formations such as photodiode, photomultiplier tube (Photomultiplier Tube), CCD (Charge Coupled Device) imageing sensor.
In said prozone phenomenon detection method of the present invention, as said, detect under the situation of oxidation current value, the said unit of obtaining comprises for example power supply and galvanometer.At first, said electrode (negative electrode and anode) is connected on the said power supply the said galvanometer of configuration between said electrode and said power supply.Then, to said electrode application voltage.Next, after said sample arrives said the 2nd test section (L2), detect said oxidation current value.Based on said oxidation current value said analytic target composition is carried out quantitatively at last.
Saidly obtaining the part that the unit can be a prozone phenomenon pick-up unit of the present invention, also can be external instrument.
Said detecting unit for example is made up of central processing unit (CPU), storer, entry terminal and outlet terminal.Said entry terminal can be enumerated out for example keyboard, touch panel.Said outlet terminal can be enumerated out for example display, printer.In addition, said detecting unit can all be configured in the prozone phenomenon pick-up unit body, also can all or part ofly be configured in the device body outside.For example also can in PC (PC), constitute detecting unit.In addition, the said various judgment standards that use in the said detecting unit can be stored in the storer and reference when detecting in advance, also can be input to through said entry terminal and carry out reference among the said CPU.Said prozone phenomenon testing result is exported through said outlet terminal.
Next, analytical equipment of the present invention is described.As previously mentioned; Analytical equipment of the present invention; It is characterized in that it comprises analytic unit and prozone phenomenon detecting unit, said analytic unit is through detecting the said mark of said marking material to the compound of said analytic target composition, said marking material and said immobilization material at the said test section of said analyte analysis tool; Thereby detect said analytic target composition, said prozone phenomenon detecting unit is said prozone phenomenon pick-up unit of the present invention.
Said analytic unit is made up of central processing unit (CPU), storer, entry terminal and outlet terminal.Said entry terminal can be enumerated out for example keyboard, touch panel.Said outlet terminal can be enumerated out for example display, printer.In addition, said detecting unit both can all be configured in the analytical equipment body, also can all or part ofly be configured in the device body outside.For example also can be in PC (PC) the component analysis unit.The testing result of said analytic target composition is exported through said outlet terminal.
Below the present invention is explained more specifically that but the present invention is not limited by these embodiment through embodiment.
Embodiment
Below record is; Preparation contains the sample of CRP with various concentration, in the immune chromatograph that uses analyte analysis tool, detects the object lesson and the object lesson of making the judgment standard that can in the prozone phenomenon that comprises said B method detects, use of the generation of prozone phenomenon.
(CRP determinand)
To use as sample by the serum of a plurality of patients and healthy person collection.In addition,, confirmed CRP concentration through the latex immunoturbidimetry in advance, be categorized as sample that normal antigen-antibody reaction takes place and the sample that produces prozone phenomenon based on its CRP concentration each serum.Particularly, with each sample of CRP concentration 0.6mg/100mL, 2.5mg/100mL and 6.5mg/100mL as the sample that normal antigen-antibody reaction takes place, with the sample more than the CRP concentration 25mg/100mL as the sample that produces prozone phenomenon.
(analyte analysis tool)
Commodity in use name " SPOTCHEMi-Line CRP " (love section comes (strain) system) is measured the analyte analysis tool of usefulness as CRP.The overview of this analyte analysis tool is as shown in Figure 4.(A) of Fig. 4 is the planimetric map of analyte analysis tool 20, and (B) of Fig. 4 is the I-I direction cut-open view of said Fig. 4 (A), and (C) of Fig. 4 is the planimetric map of in housing 31, taking in the analyte analysis chip 30 of said analyte analysis tool 20.Identically with Fig. 3 among this figure be partly with identical Reference numeral.Analyte analysis tool 20 possesses: upper substrate (not shown), infrabasal plate 21, detect with 22,2 samples of porous body the anti-CRP antibody of blue latex mark arranged with porous body 23 and 24, load marking antibody with porous body 25 and be used to absorb remain determinand absorption with porous body 26.On infrabasal plate 21, detection is arranged with porous body 22 the central range upon range of of its length direction.Detect a end (being the left side in the figure) with porous body 22 range upon range of load is arranged the marking antibody of the anti-CRP antibody of blue latex mark with porous body 25.Marking antibody has two-layer sample with porous body 23,24 with porous body 25 laminated.Absorption is arranged with porous body 26 in detection with the other end (being the right side in the figure) of porous body 22 is range upon range of.Detect with porous body 22 based on the moving direction of sample and with range upon range of have sample with the side (being the left side in the figure) of porous body 23 and 24 be the upper reaches, absorption is arranged is downstream with a side (being the right side in the figure) of porous body 26 with range upon range of, has the 1st test section (L1) and the 2nd test section (L2) that is fixed with anti-CRP antibody and the contrast that is fixed with anti-IgG antibody from the upper reaches with test section (C).The zone that will comprise the 1st test section (L1) and the 2nd test section (L2) and contrast with test section (C) is called test section 27.Then, the mode that is laminated in the various porous bodies on the infrabasal plate 21 with covering disposes upper substrate (not shown).In addition, upper substrate has through hole with sample with the corresponding position of the sample supply portion of porous body 24, in addition, expose with the 1st test section (L1), the 2nd test section (L2) and contrast with the corresponding position of test section (C).From the distance of the central portion of the center of said through hole to said the 1st test section (L1) be 30mm, to the distance of the central portion of said the 2nd test section (L2) be 33.5mm, to said contrast be 38mm with the distance of the central portion of test section (C).
(optical signalling mensuration)
In said analyte analysis tool, add each sample 5 μ L, add the back and be arranged in the reflected light determinator (trade name SPOTCHEM (registered trademark) IL, love section come (strain) system) with interior, detect the reflected light that is used to represent to have or not colour developing and colour developing degree at 1 minute.Reflected light is that the moment that is arranged at device is made as when detecting beginning (0 second) and whenever detected through 1 minute.In addition, catoptrical detection is in the moving direction of sample, test section 27 to be carried out.
To CRP concentration in the said various samples is that 1 sample of 60mg/100mL and 1 sample that CRP concentration is 6.5mg/100mL detect the detection figure that obtains from the reflected light that detects after beginning time to 10 minute respectively like (A) of Fig. 1 with (B).(A) of Fig. 1 is the result of the sample of CRP concentration 60mg/100mL, and (B) of Fig. 1 is the result of the sample of CRP concentration 6.5mg/100mL.Among two figure, the X axle is represented the coordinate of downstream from the upper reaches in the test section of said analyte analysis tool, and each scale is about 0.167mm.Among two figure, the detected value that the Y axle is represented to utilize said reflection accessory set to record (unit is counting), the more little colour developing degree of then representing of value is strong more, also is that reflectivity is low more.
Here, in the detection figure of Fig. 1 (A), the peak position of figure is positioned at the downstream of the moving direction of sample, in the prozone phenomenon of said A method detects, is judged as " having produced prozone phenomenon ".In the detection figure of Fig. 1 (B), the peak position of figure is positioned at the upstream side of the moving direction of sample, in the prozone phenomenon of said A method detects, is not judged as " having produced prozone phenomenon ".
Next, by the testing result of each sample calculate the 2nd test section, begin after 5 minutes and the reflectivity after 10 minutes from detection, calculate the increase degree of the reflectivity behind the reflectivity to 10 minute after said 5 minutes.Particularly,, prepare, at first detect contrast with near (the paddy type peak (C) of Y80～Y110) of the coordinate of test section like (A) of said Fig. 1 and (B) figure of the relation of illustrative, denotation coordination and detected value to each sample.Under the situation that can't detect this paddy type peak, think that the anti-CRP antibody of sample and blue latex mark does not normally move, and therefore can't detect, and from evaluation object, remove.Next, (Y30～Y80) detects 2 paddy type peaks near the coordinate of the 1st test section (L1) and the 2nd test section (L2).The border of the 1st test section (L1) and the 2nd test section (L2) is confirmed as at the type peak, mountain that will be positioned at the border at said 2 paddy type peaks.With the coordinate on this border as the terminal coordinate (U) of the upstream side of the 2nd test section (L2), on the other hand, will from the coordinate on said border downstream side add that the coordinate of Y20 confirms as the terminal coordinate (D) in downstream of the 2nd test section (L2).Then, connect the figure of the detected value of the terminal coordinate (D) in figure and the said downstream of detected value of the terminal coordinate (U) of said upstream side with line segment, with said line segment and the peak of paddy type figure institute area surrounded as the 2nd test section (L2) integral body.In the present embodiment, and then confirm coordinate (M), and used the result (the oblique line portions of two figure) in the zone, downstream of the 2nd test section (L2) through the mid point of said line segment.To the regional result in the downstream of the 2nd test section (L2), count value is converted into the ratio (%) from said line segment, add up to obtain reflectivity (%).In this wise each sample is obtained the downstream reflectivity (R zone, after 5 minutes of the 2nd test section (L2)
5) and the reflectivity (R after 10 minutes
10), and then calculate the reflectivity (R after 5 minutes through following formula
5) and the reflectivity (R after 10 minutes
10) poor (Δ R
5-10).
ΔR
5-10(％)＝R
5-R
10
Here, " reflectivity after 5 minutes " can use by detection and begin the value that the detected value after 5 minutes is calculated, and also for example can use " mean value of the value of being calculated by 4 minutes, 5 minutes, 6 minutes detected value ".
These results are as shown in Figure 2.This figure is the reflectivity (R of expression after 10 minutes
10) and poor (the Δ R of reflectivity
5-10) the chart of relation.In the figure, the X axle is poor (the Δ R of reflectivity
5-10), the Y axle is the reflectivity (R after 10 minutes
10), the sample of normal antigen-antibody reaction can take place in zero expression, ◆ expressing possibility produces the sample of prozone phenomenon.As shown in the drawing, possibly produce the intensive lower left quarter that is present in chart of icon of the sample (◆) of prozone phenomenon, relative therewith, the intensive upper right quarter that is present in chart of icon of the sample (zero) of normal antigen-antibody reaction can take place.Carried out being used to separate the parsing of two groups of samples, the result is as shown in the drawing, has obtained the formula of Y=-4.52x+0.75.Through with this formula as typical curve, even the unknown sample of CRP concentration, also can be by the reflectivity (R after 10 minutes
10) and poor (the Δ R of reflectivity
5-10) judge the generation have or not prozone phenomenon.
Industry is utilized possibility
As stated, according to the present invention,, can efficiently for example use the inspection of immunochromatography etc. even use existing analyte analysis tool also can detect the generation of prozone phenomenon simply.Prozone phenomenon detection method of the present invention, analytical approach, prozone phenomenon pick-up unit and analytical equipment can be applied in the fields such as clinical examination, biochemical analysis, medical research, and its purposes is unqualified, can be applied in the wide field.
Claims (14)
1. a prozone phenomenon detection method is characterized in that,
Use contain with sample in the analyte analysis tool of the material that combines of analytic target composition specificity,
Said analyte analysis tool is for disposing the analyte analysis tool of sample supply portion, reagent portion and test section downstream from the upper reaches of the moving direction of said sample on porous substrate,
The marking material that combines with said analytic target composition specificity is contained in said reagent portion,
Said test section contains the immobilization material that combines with said analytic target composition specificity,
Through the compound of said analytic target composition, said marking material and said immobilization material being detected the said mark of said marking material, thereby detect said analytic target composition at said test section,
Said method comprises at least a in following A method and the B method,
A method: at said test section, testing result is mapped, by the generation of the position probing prozone phenomenon at the peak of said figure along the moving direction of said sample;
B method:, change time and implement the detection of 2 the above marks, detect the generation of prozone phenomenon by the magnitude relationship of 2 above testing results that obtain at said test section.
2. prozone phenomenon detection method according to claim 1; It is characterized in that; This prozone phenomenon detection method comprises said B method; With reference to make in advance, the magnitude relationship of said 2 above testing results has been carried out related judgment standard with the relation of the generation of prozone phenomenon, detect the generation of said prozone phenomenon.
3. prozone phenomenon detection method according to claim 1 and 2 is characterized in that,
This prozone phenomenon detection method comprises said B method, and the magnitude relationship of said testing result is at least a in the ratio of difference and 2 above testing results of 2 above testing results.
4. according to any described prozone phenomenon detection method in the claim 1～3, it is characterized in that,
This prozone phenomenon detection method comprises said A method, when being positioned at the downstream of moving direction of sample in the peak position of said figure, being judged as and having produced prozone phenomenon.
5. prozone phenomenon detection method according to claim 4 is characterized in that,
Being judged as under the situation that produces prozone phenomenon, further carry out said B method.
6. according to any described prozone phenomenon detection method in the claim 1～5, it is characterized in that,
Said test section is 2 above test sections along the moving direction configuration of sample; In said 2 above test sections; The test section of the upstream side of the moving direction of sample is the test section that is used for the check and analysis object component, and the test section in downstream is the test section that is used to detect prozone phenomenon.
7. prozone phenomenon detection method according to claim 6 is characterized in that,
At the test section that is used for detecting prozone phenomenon, the testing result in the zone in the downstream of the moving direction through said sample detects prozone phenomenon.
8. according to any described prozone phenomenon detection method in the claim 1～7, it is characterized in that,
Said testing result is an optical signalling.
9. according to any described prozone phenomenon detection method in the claim 1～8, it is characterized in that,
Said analytic target composition is an antigen,
Said marking material and said immobilization material are marking antibody and immobilized antibody.
10. according to any described prozone phenomenon detection method in the claim 1～8, it is characterized in that,
Said analytic target composition is an antibody,
Said marking material and said immobilization material are marking antibody and immobilized antigen.
11. an analytical approach is characterized in that,
Comprise analysis procedure and prozone phenomenon and detect operation,
Said analysis procedure uses analyte analysis tool to implement,
Said analyte analysis tool is for disposing the analyte analysis tool of sample supply portion, reagent portion and test section downstream from the upper reaches of the moving direction of said sample on porous substrate,
The marking material that combines with said analytic target composition specificity is contained in said reagent portion,
Said test section contains the immobilization material that combines with said analytic target composition specificity; Through the compound of said analytic target composition, said marking material and said immobilization material being detected the said mark of said marking material at said test section; Thereby detect said analytic target composition
Said prozone phenomenon detects operation and implements through any described method in the claim 1～10.
12. analytical approach according to claim 11, said prozone phenomenon are judged to be false negative when detecting the generation that detects prozone phenomenon in the operation.
13. a prozone phenomenon pick-up unit is characterized in that,
It is used in any described prozone phenomenon detection method of claim 1～10,
The unit of obtaining that comprises the testing result that is used to obtain said test section, and
At least a in following A unit and the B unit,
The A unit: the detecting unit of the generation of the position probing prozone phenomenon at peak that get, said testing result by mapping along the moving direction of said sample,
B unit: change time and implement the detection of 2 the above marks, detect the detecting unit of the generation of prozone phenomenon by the magnitude relationship of 2 above testing results that obtain.
14. an analytical equipment is characterized in that,
It is used for claim 11 or 12 described analytical approachs,
Comprise analytic unit and prozone phenomenon detecting unit,
Said analytic unit is through detecting the said mark of said marking material to the compound of said analytic target composition, said marking material and said immobilization material at the said test section of said analyte analysis tool; Thereby detect said analytic target composition
Said prozone phenomenon detecting unit is the described prozone phenomenon pick-up unit of claim 13.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2009185298 | 2009-08-07 | ||
| JP2009-185298 | 2009-08-07 | ||
| PCT/JP2010/062066 WO2011016326A1 (en) | 2009-08-07 | 2010-07-16 | Method for detecting prozone phenomenon, analysis method, device for detecting prozone phenomenon and analyzer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102341706A true CN102341706A (en) | 2012-02-01 |
| CN102341706B CN102341706B (en) | 2014-06-11 |
Family
ID=43544224
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201080009898.9A Active CN102341706B (en) | 2009-08-07 | 2010-07-16 | Method for Detecting Prozone Phenomenon, Analysis Method, Device for Detecting Prozone Phenomenon, and Analysis Device |
Country Status (6)
| Country | Link |
|---|---|
| US (1) | US8993344B2 (en) |
| EP (1) | EP2423686B1 (en) |
| JP (1) | JP5714912B2 (en) |
| KR (1) | KR101280650B1 (en) |
| CN (1) | CN102341706B (en) |
| WO (1) | WO2011016326A1 (en) |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103376327A (en) * | 2012-04-28 | 2013-10-30 | 通用电气公司 | Method for detecting concentration of antibody or fusion protein |
| CN104714008A (en) * | 2015-02-04 | 2015-06-17 | 上海交通大学 | Immunochromatographic test strip, and making method and detection method thereof |
| CN104969069A (en) * | 2012-12-28 | 2015-10-07 | 雅培医护站股份有限公司 | Apparatus and method for identifying a hook effect and expanding the dynamic range in point of care immunoassays |
| CN107102153A (en) * | 2017-05-18 | 2017-08-29 | 天津市宝坻区人民医院 | LP(a) priority double reagent assay method in serum |
| WO2018095314A1 (en) * | 2016-11-22 | 2018-05-31 | 北京科美生物技术有限公司 | Method, system, reagent kit and system for verifying hd-hook effect sample and for performing immunoassay |
| CN108132344A (en) * | 2016-11-22 | 2018-06-08 | 博阳生物科技（上海）有限公司 | Method of immunity, for identifying the system of immunoassays and kit |
| CN108152505A (en) * | 2016-11-22 | 2018-06-12 | 博阳生物科技（上海）有限公司 | Method of immunity, for identifying the system of immunoassays and kit |
| CN108204959A (en) * | 2016-11-22 | 2018-06-26 | 博阳生物科技（上海）有限公司 | Differentiate the method for HD-HOOK effect samples and the system of the HD-HOOK effects in identification immunoassays |
| CN110114677A (en) * | 2016-12-15 | 2019-08-09 | 株式会社堀场制作所 | The determination method of the appropriateness of the test substance concentration in the concentration mensuration of immune aggregation and the sample analyzer with the processing unit for the determination method are used |
| WO2019223692A1 (en) * | 2018-05-21 | 2019-11-28 | 博阳生物科技（上海）有限公司 | Chemical luminescence analysis and measurement method, system using same, and kit |
| CN110514647A (en) * | 2018-05-21 | 2019-11-29 | 博阳生物科技（上海）有限公司 | A kind of chemiluminescence analysis measuring method and the system using this method, kit |
| CN110567900A (en) * | 2019-09-29 | 2019-12-13 | 迈克医疗电子有限公司 | method and device for judging antigen excess in sample reaction and optical detection system |
| CN116298324A (en) * | 2023-05-25 | 2023-06-23 | 武汉大学人民医院（湖北省人民医院） | Method, device, equipment and readable storage medium for detecting beta 2-microglobulin |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2880430B1 (en) | 2012-08-02 | 2018-10-10 | The Regents of The University of California | Integrated nanowire array devices for detecting and/or applying electrical signals to tissue |
| WO2014119544A1 (en) * | 2013-01-29 | 2014-08-07 | 国立大学法人三重大学 | Immunochromatographic assay method for plasma protein |
| WO2014196803A1 (en) * | 2013-06-04 | 2014-12-11 | Proteometech Inc. | A strip for rapid testing having a variable control line and a diagnosis kit using the same |
| JP2019113425A (en) * | 2017-12-25 | 2019-07-11 | Ｂｌｕｅ Ｉｎｄｕｓｔｒｉｅｓ株式会社 | Inspection cartridge and inspection device for immunochromatography assay |
| WO2020068574A1 (en) * | 2018-09-28 | 2020-04-02 | Siemens Healthcare Diagnostics Inc. | Methods for detecting hook effect(s) associated with anaylte(s) of interest during or resulting from the conductance of diagnostic assay(s) |
| CN110542660A (en) * | 2019-09-29 | 2019-12-06 | 迈克医疗电子有限公司 | method, device and detection system for detecting prozone effect in sample reaction |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0987551A2 (en) * | 1998-07-27 | 2000-03-22 | Bayer Corporation | Method for the determination of analyte concentration in a lateral flow sandwich immunoassay exhibiting high-dose hook effect |
| JP2002122599A (en) * | 2000-10-17 | 2002-04-26 | Nitto Denko Corp | Immunoassay |
| CN1473270A (en) * | 2001-08-09 | 2004-02-04 | ���µ�����ҵ��ʽ���� | Biosensors and measurement method |
| CN1576843A (en) * | 2003-07-07 | 2005-02-09 | 松下电器产业株式会社 | Immunoassay method and immunoassay system |
| JP2006250787A (en) * | 2005-03-11 | 2006-09-21 | Matsushita Electric Ind Co Ltd | Chromatography measuring device |
| CN101326440A (en) * | 2005-04-29 | 2008-12-17 | 金伯利-克拉克环球有限公司 | Assay devices having detection capabilities within the hook effect region |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06250787A (en) | 1993-02-25 | 1994-09-09 | Toshiba Corp | Display device with coordinate input function |
| JPH08278305A (en) | 1995-04-03 | 1996-10-22 | Nippon Shoji Kk | Immunological measuring instrument |
| JP3644780B2 (en) | 1996-12-25 | 2005-05-11 | 湧永製薬株式会社 | Immunological quantification equipment |
| WO2002052265A1 (en) * | 2000-12-26 | 2002-07-04 | Matsushita Electric Industrial Co., Ltd. | Specific bonding analysis method and specific bonding analysis device using it |
| EP1845373A1 (en) * | 2006-04-13 | 2007-10-17 | Olympus Life and Material Science Europa GmbH | An immunoassay method |
| AU2010236424B2 (en) * | 2009-04-15 | 2016-03-10 | Relia Biotechnologies (Shenzhen) Ltd. | Diagnostic devices and related methods |
-
2010
- 2010-07-16 KR KR1020117011009A patent/KR101280650B1/en active IP Right Grant
- 2010-07-16 CN CN201080009898.9A patent/CN102341706B/en active Active
- 2010-07-16 US US13/258,374 patent/US8993344B2/en active Active
- 2010-07-16 EP EP10806326.4A patent/EP2423686B1/en active Active
- 2010-07-16 JP JP2010547892A patent/JP5714912B2/en active Active
- 2010-07-16 WO PCT/JP2010/062066 patent/WO2011016326A1/en active Application Filing
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0987551A2 (en) * | 1998-07-27 | 2000-03-22 | Bayer Corporation | Method for the determination of analyte concentration in a lateral flow sandwich immunoassay exhibiting high-dose hook effect |
| JP2002122599A (en) * | 2000-10-17 | 2002-04-26 | Nitto Denko Corp | Immunoassay |
| CN1473270A (en) * | 2001-08-09 | 2004-02-04 | ���µ�����ҵ��ʽ���� | Biosensors and measurement method |
| CN1576843A (en) * | 2003-07-07 | 2005-02-09 | 松下电器产业株式会社 | Immunoassay method and immunoassay system |
| JP2006250787A (en) * | 2005-03-11 | 2006-09-21 | Matsushita Electric Ind Co Ltd | Chromatography measuring device |
| CN101326440A (en) * | 2005-04-29 | 2008-12-17 | 金伯利-克拉克环球有限公司 | Assay devices having detection capabilities within the hook effect region |
Cited By (32)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103376327A (en) * | 2012-04-28 | 2013-10-30 | 通用电气公司 | Method for detecting concentration of antibody or fusion protein |
| CN104969069B (en) * | 2012-12-28 | 2017-09-12 | 雅培医护站股份有限公司 | For the apparatus and method for the dynamic range for identifying hook effect and expansion point of care immunoassays |
| CN104969069A (en) * | 2012-12-28 | 2015-10-07 | 雅培医护站股份有限公司 | Apparatus and method for identifying a hook effect and expanding the dynamic range in point of care immunoassays |
| US9671398B2 (en) | 2012-12-28 | 2017-06-06 | Abbott Point Of Care Inc. | Apparatus and method for identifying a hook effect and expanding the dynamic range in point of care immunoassays |
| CN104714008A (en) * | 2015-02-04 | 2015-06-17 | 上海交通大学 | Immunochromatographic test strip, and making method and detection method thereof |
| CN109470866A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| CN109470863A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| CN108132344A (en) * | 2016-11-22 | 2018-06-08 | 博阳生物科技（上海）有限公司 | Method of immunity, for identifying the system of immunoassays and kit |
| CN108152505A (en) * | 2016-11-22 | 2018-06-12 | 博阳生物科技（上海）有限公司 | Method of immunity, for identifying the system of immunoassays and kit |
| CN108204959A (en) * | 2016-11-22 | 2018-06-26 | 博阳生物科技（上海）有限公司 | Differentiate the method for HD-HOOK effect samples and the system of the HD-HOOK effects in identification immunoassays |
| CN109470861A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| CN109470871A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| CN109470870A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| CN109470862A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| CN109470860A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| CN109470868A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| CN109470868B (en) * | 2016-11-22 | 2022-04-15 | 科美博阳诊断技术(上海)有限公司 | Immunoassay method and system for hepatitis C virus antibody |
| CN109470856A (en) * | 2016-11-22 | 2019-03-15 | 博阳生物科技（上海）有限公司 | Method of immunity, the system for identifying immunoassays and kit |
| WO2018095314A1 (en) * | 2016-11-22 | 2018-05-31 | 北京科美生物技术有限公司 | Method, system, reagent kit and system for verifying hd-hook effect sample and for performing immunoassay |
| CN109470870B (en) * | 2016-11-22 | 2022-04-15 | 科美博阳诊断技术(上海)有限公司 | Immunoassay method |
| CN109470863B (en) * | 2016-11-22 | 2021-12-17 | 科美博阳诊断技术(上海)有限公司 | Immunoassay method, system and kit for identifying immunoassay |
| CN109470860B (en) * | 2016-11-22 | 2021-12-10 | 科美博阳诊断技术(上海)有限公司 | Immunoassay method, system and kit for identifying immunoassay |
| CN109470862B (en) * | 2016-11-22 | 2021-12-03 | 科美博阳诊断技术(上海)有限公司 | Immunoassay method, system and kit for identifying immunoassay |
| CN110114677A (en) * | 2016-12-15 | 2019-08-09 | 株式会社堀场制作所 | The determination method of the appropriateness of the test substance concentration in the concentration mensuration of immune aggregation and the sample analyzer with the processing unit for the determination method are used |
| CN110114677B (en) * | 2016-12-15 | 2023-06-20 | 株式会社堀场制作所 | Method for determining suitability of concentration of test substance in concentration measurement using immune coagulation reaction, and sample analyzer having processing unit for the determination method |
| CN107102153A (en) * | 2017-05-18 | 2017-08-29 | 天津市宝坻区人民医院 | LP(a) priority double reagent assay method in serum |
| CN110514647A (en) * | 2018-05-21 | 2019-11-29 | 博阳生物科技（上海）有限公司 | A kind of chemiluminescence analysis measuring method and the system using this method, kit |
| WO2019223692A1 (en) * | 2018-05-21 | 2019-11-28 | 博阳生物科技（上海）有限公司 | Chemical luminescence analysis and measurement method, system using same, and kit |
| CN110514647B (en) * | 2018-05-21 | 2023-01-20 | 科美博阳诊断技术(上海)有限公司 | Chemiluminescence analysis determination method, system and kit using same |
| CN110567900A (en) * | 2019-09-29 | 2019-12-13 | 迈克医疗电子有限公司 | method and device for judging antigen excess in sample reaction and optical detection system |
| CN110567900B (en) * | 2019-09-29 | 2022-07-05 | 迈克医疗电子有限公司 | Method and device for judging antigen excess in sample reaction and optical detection system |
| CN116298324A (en) * | 2023-05-25 | 2023-06-23 | 武汉大学人民医院（湖北省人民医院） | Method, device, equipment and readable storage medium for detecting beta 2-microglobulin |
Also Published As
| Publication number | Publication date |
|---|---|
| JPWO2011016326A1 (en) | 2013-01-10 |
| EP2423686A4 (en) | 2013-01-02 |
| CN102341706B (en) | 2014-06-11 |
| EP2423686B1 (en) | 2015-09-02 |
| US20120015450A1 (en) | 2012-01-19 |
| EP2423686A1 (en) | 2012-02-29 |
| JP5714912B2 (en) | 2015-05-07 |
| KR20110084253A (en) | 2011-07-21 |
| US8993344B2 (en) | 2015-03-31 |
| WO2011016326A1 (en) | 2011-02-10 |
| KR101280650B1 (en) | 2013-07-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102341706B (en) | Method for Detecting Prozone Phenomenon, Analysis Method, Device for Detecting Prozone Phenomenon, and Analysis Device | |
| Shah et al. | Enzyme-linked immunosorbent assay (ELISA): the basics | |
| CN106872420B (en) | Kit and method for time-resolved fluorescence quantitative detection of microalbuminuria | |
| RU2594066C2 (en) | Enzyme immunoassay | |
| Rivnak et al. | A fully-automated, six-plex single molecule immunoassay for measuring cytokines in blood | |
| US7790439B2 (en) | Biosensor and measurement method | |
| JP3458860B2 (en) | Sensor device for sandwich analysis | |
| US8956823B2 (en) | Anti-antibody reagent | |
| CN104081207A (en) | Integrated test device for optical and electrochemical assays | |
| CN104081210A (en) | Optical assay device with pneumatic sample actuation | |
| CN105403698A (en) | Diagnostic test kits employing an internal calibration system | |
| Lin et al. | Washing-free centrifugal microchip fluorescence immunoassay for rapid and point-of-care detection of protein | |
| Abate et al. | Distance-based paper/PMMA integrated ELISA-chip for quantitative detection of immunoglobulin G | |
| JPWO2003029822A1 (en) | Specific binding analyzer and specific binding analysis method | |
| US20030119030A1 (en) | Immunoassay diagnostic probe and a method for use thereof | |
| KR20160134796A (en) | Glycated protein assay | |
| KR20190058357A (en) | A wide range of in-vitro diagnostic kit capable of quantitating target substance in high concentration specimen | |
| Shin et al. | Hook effect detection and detection-range-controllable one-step immunosensor for inflammation monitoring | |
| JP2019113425A (en) | Inspection cartridge and inspection device for immunochromatography assay | |
| JP2010091308A (en) | Method for diagnosing prostate carcinoma by lectin absorbing method and prostate carcinoma determining kit | |
| CN113113079B (en) | Method for identifying hook effect in quantitative immunochromatography test | |
| CN204287192U (en) | Myeloperoxidase immunochromatographiassay assay quantitative detection test paper | |
| Zhang et al. | An automated, rapid fluorescent immunoassay to quantify serum soluble programmed death-1 (PD-1) protein using testing-on-a-probe biosensors | |
| CN103185779B (en) | Reagent device for detecting antinuclear antibody and method thereof | |
| CN114814243B (en) | Quantitative detection kit and method applied to protein antigen |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |